Motor nerve terminal destruction and regeneration following anti-ganglioside antibody and complement-mediated injury: An in and ex vivo imaging study in the mouse
Motor nerve terminal destruction and regeneration following anti-ganglioside antibody and complement-mediated injury: An in and ex vivo imaging study in the mouse
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DOI：
10.1016/j.expneurol.2011.12.010
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发表时间：
2012-02-01
影响因子：
5.3
通讯作者：
Willison, Hugh J.
中科院分区：
文献类型：
--
作者：
Rupp, Angie;Morrison, Ian;Willison, Hugh J.
在人类周围神经疾病格林-巴利综合征 (GBS) 的小鼠模型中，神经肌肉接头 (NMJ) 的神经和神经胶质成分已被确定为抗神经节苷脂抗体 (Ab) 结合和补体介导损伤的潜在位点。一些患有急性运动轴突神经病（AMAN）形式的 CBS 的患者从瘫痪中恢复得非常快；有人提出，在这些病例中，损伤仅限于能够在很短的时间内再生的远端运动轴突和神经末梢（NT）。为了检验这一假设，对轴突（CFP）和雪旺细胞（GFP）表达胞质荧光蛋白的小鼠（n = 45）的腹侧颈部肌肉进行单次局部应用抗神经节苷脂抗体，然后使用补体源。 A 组 (n = 15) 接受选择性结合 NT 的抗体，B 组 (n = 15) 接受同时结合 NT 和突触周围雪旺细胞 (pSC) 的抗体，C 组（对照动物；n = 15）仅接受补体。通过体内成像记录损伤的演变，安乐死后，立即或在再生 1、2、3 或 5 天后（每组 n = 3）对肌肉进行离体定量和定性重新成像。在补体应用 15 分钟内，可以看到覆盖 NMJ 的 CFP 迅速损失；在A组中，GFP信号保持不变，而在B组中，GFP信号也丢失。在 C 组中，没有观察到 CFP 或 GFP 的变化。 24小时时，A组中6%的浅表NMJ和B组中12%的NMJ表现出CFP。在两组中，CFP 在接下来的五天内恢复（A 组：93.5%，B 组：94%；p = 0.739），B 组中 CFP 恢复之前，NMJ 上覆盖的 GFP 阳性细胞返回。辅助研究显示，NMJ 处 CFP 的丧失与 NT 神经丝免疫反应性的丧失相关，并且 NMJ 处 CFP 的恢复伴随着神经丝的回归。在超微结构研究中，受损的 NT 具有电子透明性，并表现出线粒体受损、细丝损失和突触小泡损失。再生五天后对肌肉的检查揭示了生理性 NT 特征。上述结果表明，在单次抗神经节苷脂抗体介导和补体介导的攻击后，无论 NMJ 上是否有健康且成熟的突触周围雪旺细胞，小鼠 NT 都能够非常迅速地恢复其结构和轴突完整性。该数据支持这样一种观点，即在一些 AMAN 临床病例中可能存在一种等效的机制来解释快速恢复的原因。 (C) 2011 年，爱思唯尔公司出版。
Both the neural and glial components of the neuromuscular junction (NMJ) have been identified as potential sites for anti-ganglioside antibody (Ab) binding and complement-mediated injury in murine models for the human peripheral nerve disorder Guillain-Barre syndrome (GBS). Some patients suffering from the acute motor axonal neuropathy (AMAN) forms of CBS recover very rapidly from paralysis; it has been proposed that in these cases the injury was restricted to the distal motor axons and nerve terminals (NTs) which are able to regenerate over a very short time-frame.To test this hypothesis, the ventral neck muscles of mice (n = 45) expressing cytosolic fluorescent proteins in their axons (CFP) and Schwann cells (GFP) were subjected to a single topical application of anti-ganglioside Ab followed by a source of complement. Group A (n = 15) received Ab that selectively bound to the NTs, group B (n = 15) received Abs that bound both to the NTs and the perisynaptic Schwann cells (pSCs) and group C (control animals; n = 15) only received complement. Evolution of the injury was documented by in vivo imaging, and following euthanasia the muscles were reimaged ex vivo both quantitatively and qualitatively, either immediately, or after 1, 2, 3 or 5 days of regeneration (each n = 3 per group).Within 15 minutes of complement application, a rapid loss of CFP overlying the NMJ could be seen; in group A, the GFP signal remained unchanged, whereas in group B the GFP signal was also lost. In group C no changes to either CFP or GFP were observed. At 24 h, 6% of the superficial NMJs in group A and 12% of the NMJs in group B exhibited CFP. In both groups, CFP returned within the next five days (group A: 93.5%, group B: 94%; p = 0.739), with the recovery of CFP being preceded by a return of GFP-positive cells overlying the NMJ in group B. Auxiliary investigations revealed that the loss of CFP at the NMJ correlated with a loss of NT neurofilament immuno-reactivity and a return of CFP at the NMJ was accompanied by a return of neurofilament. In ultrastructural investigations, injured NTs were electron lucent and exhibited damaged mitochondria, a loss of filaments and a loss of synaptic vesicles. The examination of muscles after five days of regeneration revealed physiological NT-profiles.The results described above indicate that following a single anti-ganglioside Ab-mediated and complement-mediated attack, independent of whether there are healthy and mature perisynaptic Schwann cells overlying the NMJ, the murine NT is capable of recovering both its architectural and axolemmal integrity very rapidly. This data supports the notion that an equivalent mechanism may account for the rapid recovery seen in some clinical cases of AMAN. (C) 2011 Published by Elsevier Inc.